title: Model peptides provide new insights into the role of histidine residues as potential ligands in human cellular copper acquisition via Ctr1.
abstract: Cellular acquisition of copper in eukaryotes is primarily accomplished through the Ctr family of copper transport proteins. In both humans and yeast, methionine-rich "Mets" motifs in the amino-terminal extracellular domain of Ctr1 are thought to be responsible for recruitment of copper at the cell surface. Unlike yeast, mammalian Ctr1 also contains extracellular histidine-rich motifs, although a role for these regions in copper uptake has not been explored in detail. Herein, synthetic model peptides containing the first 14 residues of the extracellular domain of human Ctr1 (MDHSHHMGMSYMDS) have been prepared and evaluated for their apparent binding affinity to both Cu(I) and Cu(II). These studies reveal a high affinity Cu(II) binding site (log K = 11.0 ± 0.3 at pH 7.4) at the amino-terminus of the peptide as well as a high affinity Cu(I) site (log K = 10.2 ± 0.2 at pH 7.4) that utilizes adjacent HH residues along with an additional His or Met ligand. These model studies suggest that the histidine domains may play a direct role in copper acquisition from serum copper-binding proteins and in facilitating the reduction of Cu(II) to the active Ctr1 substrate, Cu(I). We tested this hypothesis by expressing a Ctr1 mutant lacking only extracellular histidine residues in Ctr1-knockout mouse embryonic fibroblasts. Results from live cell studies support the hypothesis that extracellular amino-terminal His residues directly participate in the copper transport function of Ctr1.
authors.affiliation: Department of Chemistry, Duke University, Durham, North Carolina 27708, USA.
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Haas
authors.forename: Allison B
authors.initials: AB
authors.lastname: Putterman
authors.forename: Daniel R
authors.initials: DR
authors.lastname: White
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: Thiele
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Franz
mesh_terms: D000818:Animals; D027682:Cation Transport Proteins / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D002460:Cell Line; D003300:Copper / Q000737:chemistry / Q000378:metabolism*; D000079923:Copper Transporter 1; D056890:Eukaryota / Q000737:chemistry / Q000378:metabolism*; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D006639:Histidine / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D051379:Mice; D018345:Mice, Knockout; D008954:Models, Biological; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D027682:Cation Transport Proteins; D000079923:Copper Transporter 1; D008024:Ligands; D010455:Peptides; C490079:SLC31A1 protein, human; C490077:Slc31a1 protein, mouse; D006639:Histidine; D003300:Copper